Acta Toxicol. Argent.

(2013) 21 (2): 93-101

Acute silver toxicity to Cnesterodon decemmaculatus (Pisces: Poeciliidae)

in a river with extreme water-quality characteristics
Toxicidad aguda de la plata sobre Cnesterodon decemmaculatus (Pisces: Poeciliidae)
en un ro con caractersticas extremas de calidad de agua
Casares, Mara Victoria1*; de Cabo, Laura I.1; Seoane, Rafael2,3; Natale, Oscar2
Bernardino Rivadavia National Museum of Natural History, Avenida Angel Gallardo 470 (C1405DJR), Buenos Aires, Argentina. Tel.
54 011 4982 4494; fax: 54 011 4982 0306. 2National Water Institute, Autopista Ezeiza-Cauelas, Tramo Jorge Newbery km 1.62
(1802) Ezeiza, Buenos Aires, Argentina. 3Faculty of Engineering, University of Buenos Aires, Avenida Las Heras 2214 (C1127AAR),
Buenos Aires, Argentina.
*mvc251@hotmail.com
1

Recibido: 12 de abril de 2013

Aceptado: 13 de agosto de 2013

Abstract. A 96 h acute silver toxicity test was performed in order to determine silver toxicity (LC50) to a local fish species (Cnesterodon decemmaculatus) in a river with extreme water-quality characteristics (Pilcomayo River, South America) and evaluate a
cross-fish-species extrapolation of the Biotic Ligand Model. The dissolved silver concentrations tested were 0.095, 0.148, 0.175
and 0.285 mg Ag L1. The 96 h Ag LC50 calculated for C. decemmaculatus was 0.14 mg L1 (0.18 - 0.10) and the value predicted
by BLM for Pimephales promelas was 0.051 mg Ag L1. Test water elevated hardness may have exerted some protective effect.
High mean water pH may have exerted a major protective effect by reducing silver free ion form and causing silver precipitation.
The mortality pattern observed in this toxicity test may lend some support to a relationship between gill silver accumulation and
mortality. A cross-fish-species extrapolation of Ag BLM for P. promelas was not valid in Pilcomayo River water and experimental
conditions of this toxicity test.
Keywords: Silver; Cnesterodon decemmaculatus; Biotic Ligand Model; Extrapolation.
Resumen. Con el objeto de determinar la toxicidad de la plata en un pez nativo (Cnesterodon decemmaculatus), se llev a cabo
un ensayo esttico de toxicidad aguda a 96 horas en un agua natural con caractersticas de calidad de agua, extremas (ro Pilcomayo, Sudamrica). Asimismo, se evalu una posible extrapolacin inter-especie del Modelo del Ligando Bitico en el agua
experimental. La concentracin inicial de plata en solucin en los distintos tratamientos fue de 0,095; 0,148; 0,175 y 0,285 mg Ag
L1. La CL50 a las 96 horas calculada para C. decemmaculatus fue de 0,14 (0.18 - 0.10) mg Ag L1 y el valor predicho por el BLM
para Pimephales promelas fue de 0,051 mg Ag L1. La elevada dureza del agua experimental pudo haber tenido algn efecto protector frente a la toxicidad de la plata. El valor medio de pH del ensayo fue elevado y posiblemente tuvo un gran efecto protector
por reduccin de la forma inica libre y precipitacin del metal. El patrn de mortalidad observado en este ensayo de toxicidad
apoyara la relacin causa-efecto entre acumulacin de plata en las branquias y mortalidad. La extrapolacin inter-especie del
BLM para P. promelas no result vlida en el agua del ro Pilcomayo y en las condiciones experimentales de este ensayo.
Palabras clave: Plata; Cnesterodon decemmaculatus; Modelo de Ligando Bitico; Extrapolacin.

INTRODUCTION
Silver, as many other heavy metals enters the
aquatic ecosystems as a by-product of many
industrial and mining processes. Silver ion is
one of the most toxic forms of a heavy metal,
surpassed only by mercury and thus, it has
been assigned to the highest toxicity class,
together with cadmium, chromium (VI), copper and mercury (Ratte 1999). The primary
effect of silver is on sodium and chloride uptake and efflux. When present as silver ni-

trate, silver is one of the most acutely toxic

metals to freshwater rainbow trout (Oncorhynchus mykiss) with 96-h LC50 values in the
range of 6.513 g L1 in freshwaters that are
generally low in organic matter content. Silver
nitrate is highly toxic because it readily dissociates in water to yield ionic silver (Ag+), the
most acutely toxic species of silver. Silver toxicity occurs in freshwater fishes as a result of
Ag+ disabling first the carbonic anhydrase and
later the enzyme Na+/K+ adenosinetriphospha-

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Acta Toxicol. Argent. (2013) 21 (2): 93-101

tase (ATPase) at the gill, leading to a net loss of

sodium and chloride (Morgan et al. 2004). As
a result, blood volume decreases and blood
viscosity and pressure increase leading to fish
mortality (Kennedy 2011).
The Biotic Ligand Model (BLM) (Di Toro et al.
2001) has been proposed as a tool to evaluate quantitatively the manner in which water
chemistry affects the speciation and biological
availability of metals in aquatic systems (Paquin et al. 2002). This model was developed
using geochemical equilibrium principles,
and provides site-specific toxicity predictions
(Clifford and McGeer 2009). It is implicitly assumed that BLMs can be extrapolated within
taxonomically similar groups, i.e., that BLMs
developed for P. promelas can be applied to
ecotoxicity data for other fish species, that
BLMs for D. magna and C. dubia can be applied to ecotoxicity data for other invertebrates
(Schlekat et al. 2010). The basis for a crossspecies extrapolation is the assumption that
the parameters, which describe interactions
between cations (notably calcium, magnesium
and protons), the toxic free metal ion (M n+) and
the biotic ligand are similar across organisms,
and that only intrinsic sensitivity varies among
species (Schlekat et al. 2010).
A variety of water chemistry factors can protect against metal binding to the sites of action of toxicity either by cationic competition
(sodium and protons in the case of silver), or
by anionic complexation (e.g., hydroxide, (bi)
carbonate, chloride, thiosulfate, sulfide, and
most importantly dissolved organic matter),
thereby preventing it binding to the toxicity
sites (Wood 2008). One of the first and most
well recognized of the modifying factors for
metals is water hardness. However, it appears
to be less important for silver toxicity (Kennedy 2011). Alkalinity, on the other hand, affects
metal ionic species in water solution through
their complexation with carbonates (Erickson
et al. 1996; Lauren and McDonald 1986) and
dissolved organic matter forms complexes
with metals, which reduces the free form in the
water, and therefore the amount of ionic metal
available to bind to the gill sites (Matsuo et al.
2004).
Pilcomayo River water is characterized by
high concentration of sulphates, chloride, calcium, magnesium, and total suspended solids.
Moreover, most of the major ions mean concentration in Pilcomayo River water is at the
95th percentile compared to the 60 largest riv-

ers in the world (data in Gaillardet et al. 1999).

Toxic waste spills containing high concentrations of arsenic and heavy metals are released
daily from the mining district of Potos, in Bolivia into Pilcomayo River waters. Mining of
the Cerro Rico de Potos ores began in 1545
however, since the introduction of the Crushing-Grinding-Flotation method of mineral processing (1985) the chronic contamination of
the Pilcomayo River water and sediments has
increased steadily (Smolders et al. 2003).
Cnesterodon decemmaculatus (Pisces: Poeciliidae; Jenyns 1842) is an endemic member of
the fish family Poeciliidae with extensive distribution in neotropical America. The species
attains high densities in a large variety of water bodies within the entire La Plata River and
other South American basins. Cnesterodon
decemmaculatus is a small, viviparous, microomnivorous, benthic-pelagic, non-migratory
fish. This species, highly tolerant to extreme
environmental conditions, is additionally easy
to handle and breed under laboratory conditions (Menni 2004). Thereby, several authors
have used C. decemmaculatus in bioassays
(de la Torre et al. 1997, 2002, 2005; Ferrari et
al. 1998; Gmez et al. 1998).
Pimephales promelas (Pisces: Cyprinidae;
Rafinesque 1820), one of the fish species for
which BLM has been developed, is a temperate, holartic fresh water fish. As well as C. decemmaculatus, is quite tolerant to turbid, lowoxygenated water bodies, and can be found
in muddy ponds and streams and in small rivers (http://www.fishbase.org/summary/Pimephales-promelas.html). There is one previous
work on the toxicity of metals to C. decemmaculatus in Pilcomayo River water (Casares
et al. 2012). The authors found that a crossfish-species extrapolation of the Cu-BLM was
valid within the Pilcomayo River water quality
characteristics and experimental conditions
of their toxicity test. With respect to Ag BLM,
more work with a wider range of species will
be required to improve its predictive capability
(Bianchini et al. 2002).
The aim of this study was to evaluate a crossfish-species extrapolation of the silver BLM
in a river with extrem water-quality characteristics (Pilcomayo River water). In order to
achieve this aim: (a) silver toxicity (96-h Ag
LC50) to C. decemmaculatus was assessed in
Pilcomayo River water, (b) BLM, version 2.2.3
was applied to predict acute silver toxicity to P.
promelas (Ag LC50) under Pilcomayo River wa-

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Acta Toxicol. Argent. (2013) 21 (2): 93-101

ter quality characteristics and (c) the predicted

Ag LC50 value for P. promelas was compared
to the calculated for C. decemmaculatus.

waters are located in Bolivia along the eastern flank of the central Andes at an elevation
of approximately 5,200 m (Figure 1). The river
flows in a southeasterly direction until reaching
the Chaco Plains along Bolivia's southern border with Argentina. Its total length is 2,426 km
and its basin covers an area of approximately
288,360 km2 (http://www.pilcomayo.net/web/).

MATERIALS AND METHODS

Study area
The Pilcomayo River in South America is a tributary to the large La Plata system. Its head-

Figure 1. Map of Pilcomayo River basin with the water sampling location (Misin La Paz, Argentina)

Water sampling and chemical analysis

Discrete water samples for chemical analysis were taken 10 cm below the water surface
and in triplicate from the navigation channel,
left and right shore of Pilcomayo River in Misin La Paz International bridge (22 22 45 S
62 31 08 W; 254 meters over sea level) in
September 2009 (Figure 1). Water discharge
(Q), pH and water temperature (T) were determined in situ. Dissolved concentrations of calcium (Ca), magnesium (Mg), chloride (Cl), potassium (K), sodium (Na), sulphate (SO4), hardness (Hard), alkalinity (Alk), dissolved organic
carbon (DOC), total suspended solids (TSS),

total dissolved solids (TDS), total (T. Ag) and

dissolved silver (D. Ag) concentrations were
determined using Standard Methods test protocols (APHA, AWWA, WPCF 1992).
Toxicity test
Water for the toxicity test was collected in prerinsed 10-L polypropylene containers. Samples were immediately placed into coolers and
transported by plane to the laboratory. Later,
water was centrifuged (2000 rpm during 15
minutes) and filtered through 47 mm, 0,45m
pore glass-fibre filters (Whatman GF/C). Dissolved silver background concentration in Pil-

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Acta Toxicol. Argent. (2013) 21 (2): 93-101

comayo River water was below the method

detection limit (ND).
Juvenile C. decemmaculatus were collected
from a small pond, located in Reserva Natural
Los Robles, Buenos Aires Province, Argentina (main chemical and physical parameters in
Casares et al. 2012). Fish were kept at temperatures ranging from 20 to 24C and pH ranging from pH 7.1 to 7.5 in an aquarium supplied
with a continuous flow of aerated de-chlorinated tap water for 30 days. During this period
and posterior laboratory and test water (centrifuged and filtered Pilcomayo River water) acclimation the fish were fed with a daily ration
of commercial fish food. Acclimation to test
water was performed by adding small quantities of test water to the aquarium until most
of the water volume corresponded to test water. One day before and during the experiment,
fish were not fed.
Toxicity effect of silver on fish was tested in
static systems (4L glass aquaria) with continuous artificial aeration, constant environmental
temperature and natural laboratory photoperiod. Test water volume in each aquarium was
2L. Test silver concentrations were attained by
spiking from stock solutions of 200 mg Ag L-1.
The reagent-grade toxicants used to perform
the stock solutions was AgNO3 (Merck analytical grade).
To define the range of silver concentrations to
be employed in the bioassay a nominal concentration of 1 mg Ag L-1 was tested in an
aquarium with 2L volume of Pilcomayo River
water and 10 acclimated specimens of juvenile
C. decemmacuatus for 96 h. Subsequently,
the experimental design applied included different metal concentrations or treatments (T)
with one control group (kept in test water and
without metal addition). Fish (not sexed) taken
from the acclimation tank were randomly distributed in the different experimental aquaria.
Mean standard length of the specimens selected was 13.8 1.2mm and each aquarium
contained 10 specimens. Metal concentration
in the experimental aquaria was adjusted prior
to the fish transfer. Survival was registered four
times a day during 96 h. Water pH, conductivity and dissolved oxygen were measured with
portable probes from HANNA (HANNA instruments, Inc. Woonsocket, RI, USA) daily. At the
beginning of the toxicity test, water samples
were collected into polypropylene conical
tubes and acidified to pH<2 with concentrated
nitric acid (reagent-grade) for metal analysis.

Silver in samples was measured by graphite

furnace atomic absorption spectrophotometry
Perkin Elmer AAnalyst 800 (Perkin Elmer, Inc.
Waltham, MA, USA). Method detection limit
was 0.008 mg L-1.
LC50 calculations
The median lethal concentration (LC50) at 24,
48, 72 and 96 hours (24-h LC50, 48-h LC50,
72-h LC50, 96-h LC50) were calculated using
PROBIT method (Finney 1978) and the statistical program StatPlus (Analyst Soft Inc.).
Version 2.2.3 of the BLM Windows Interface
(available at http://www.hydroqual.com/wr_
blm.html) was run in order to predict acute
silver toxicity to P. promelas (toxicity mode).
Pilcomayo River water quality parameters employed to run the BLM were temperature, pH,
dissolved organic carbon, calcium, magnesium, sodium, potassium, sulphate, chloride
and alkalinity.
Metal speciation
BLM (speciation mode) was also used to assess chemical speciation of the measured silver concentrations.
RESULTS
Water quality parameters of Pilcomayo River
test water are shown in table 1.
Table 1. Test water main physicochemical
parameters (data provided by Subsecretara de
Recursos Hdricos - Argentina). ND: not detected.
Parameter
Q m3 s-1 19.3
pH UpH

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7.7

Ca

mg L-1 119

Mg

mg L-1 46.2

Cl

mg L-1 208

Na

mg L-1 143.9

mg L-1 12

SO4

mg L-1 364

DOC

mg L-1 3

Alk

mg L-1 CaCO3

142

Hard

mg L-1 CaCO3

485.16

T. Ag

mg L-1 ND

D. Ag

mg L-1 ND

TSS

mg L-1 546

TDS

mg L-1 945

Acta Toxicol. Argent. (2013) 21 (2): 93-101

Toxicity test mean water pH and temperature

were 8.3 0.2 and 25.1 1.2 C, respectively.
Within the first few hours from the beginning of
the toxicity test, a precipitate was observed. No
mortality was observed in the control group. Initial exposure silver concentrations were 0.095
(T1), 0.148 (T2), 0.175 (T3) and 0.285 mg Ag L-1
(T4). Highest mortality was observed within the

first five hours in all treatments.

The median lethal concentrations (LC50, mg L-1)
at 24, 48, 72 and 96 h (24-h LC50, 48-h LC50,
72-h LC50, 96-h LC50) with their corresponding
confidence intervals (quoted) calculated using
PROBIT method and the 96-h Ag LC50 predicted by BLM for P. promelas in the toxicity
test water are shown in table 2.

Table 2. The LC50 at 24, 48, 72 and 96 hours (24-h LC50, 48-h LC50, 72-h LC50, 96-h
LC50) with their respective confidence intervals calculated for C. decemmaculatus
and the predicted 96-h LC50 by BLM for P. promelas in the test water.
LC50 (mg Ag L-1)
24-h

48-h

72-h

96-h

BLM

0.21 (0.15-0.33)

0.17 (0.12-0.22)

0.15 (0.11-0.19)

Figure 2 shows calculated silver toxicity to C.

decemmaculatus compared with predicted silver toxicity (LC50, in g Ag L-1) using the Biotic
Ligand Model. As it can be observed BLM prediction was not accurate within a factor of 2.
BLM speciation output can be observed in figure 3. Silver speciation in the four treatments
was practically the same with 60% of the silver
as AgCl and 30% as AgCl2. The free ion form
represented the highest contribution (6%)
amongst the remaining species group.

0.14 (0.10-0.18)

0.051

of water hardness on copper toxicity have

been well documented; whereas, its protective
effect on silver toxicity seems to be not clear.
For Paquin et al. (2002), when competing with
silver at the biotic ligand, calcium tends to
be of less importance than it appears to be
for other metals. Bury et al. (1998) found that
an increment on calcium concentration from
2 to 80 mg L-1 had a small influence on the
96-h LC50 for rainbow trout and P. promelas.
These authors reported a 96-h Ag LC50 for juvenile P. promelas of 0.008 mg L-1. Karen et
al. (1999) reported that dissolved organic carbon was more important than hardness for
predicting the toxicity of ionic silver in natural

DISCUSSION
Pilcomayo River water characterizes by its
high water hardness. The ameliorative effects

100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
T1

T2
AgCl

Figure 2. Calculated silver toxicity to C. decemmaculatus

compared with predicted silver toxicity (LC50, in g Ag
L-1) using the Biotic Ligand Model developed for P.
promelas. The thicker line represents a 1:1 relationship.
The thinner lines represents predictions within a factor
of 2. The error bars represent 95% confidence intervals.

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Treatments
Ag(Cl)2

T3

T4

Remaing species

Figure 3. BLM speciation output for each treatment.

Silver species are expressed as percentages of
total dissolved silver concentration. Remaining
species summeraizes the contribution of the
following species: free ion, AgCl3, AgCl4, AgNO3,
AgSH, Ag(SH)2, Torg Ag, AgOH and Ag(OH)2.

Acta Toxicol. Argent. (2013) 21 (2): 93-101

waters to O. mykiss, P. promelas, and D. magna. On the other hand, Erickson et al. (1998)
reported a substantial decrease in silver toxicity to P. promelas (96-h LC50 increase from
0.005 to 0.012 mg L-1) when water hardness
was increased from 48 to 249 mg L-1 CaCO3.
Bielmyer et al. (2007) found that C. dubia and
P. promelas were less sensitive to silver in waters with a combination of higher hardness
and dissolved organic carbon. In water with a
hardness of 225 mg L-1 CaCO3, these authors
reported a 96-h LC50 for 7-days old P. promelas of 0.021 (0.019-0.023) mg L-1. Despite the
opposite results reported, higher 96-h LC50
estimated for C. decemmaculatus may reflect
some protective effect exerted by Pilcomayo
River water elevated hardness.
High levels of sodium are thought to inhibit silver accumulation through competition; however, Bury and Wood (1999) found that one-third
of the silver uptake continued in presence of a
sodium channel and ATPase blockers or high
levels of sodium in water. Erickson et al. (1998)
reported no significant effects on silver toxicity to P. promelas when 2 meq L-1 of sodium
sulphate were added to test water. This means
that alternatively or additionally, multiple pathways exist for apical silver uptake at the gill
cells. If multiple silver pathways are also present in C. decemmaculatus, the high levels of
sodium in Pilcomayo River water did not exert
a protective effect against silver toxicity.
Pilcomayo River dissolved organic matter levels are relatively low. Increased levels of organic carbon are expected to affect silver bioavailability through complexation. Bury et al. (1998)
observed that an increase of 0.3 to 5.8 mg L-1
in dissolved organic carbon reduced acute silver toxicity to rainbow trout and P. promelas.
Erickson et al. (1998) reported that increased
levels of dissolved organic carbon (from 1 to
11 mg C L-1) significantly increased Ag LC50 for
30-day-old P. promelas. However, Rose-Janes
and Playle (2000) highlighted the strong binding of Ag+ to trout gills and the relatively weaker binding of silver to dissolved organic matter.
In the present contribution, dissolved organic
carbon did not exert an important effect on silver speciation, since the predicted percentage
of silver bound to dissolve organic matter was
very low (0.52 %).
A net loss of chloride is also observed during
silver exposure. The protective effect of high
levels of chloride in water has been shown to
be different among species. Some species

rely on branchial chloride uptake, which, in

some species, is not inhibited by silver exposure, and others depend on multiple chloride
uptake pathways. Apparently, chloride uptake
pathways in P. promelas are not inhibited by
silver exposure (Bielmyer et al. 2008). Given
that chloride uptake pathways in C. decemmaculatus are unknown, we cannot suggest
that high chloride concentration in Pilcomayo
River water could have exerted a protective
effect against silver toxicity. External chloride may also exert a protective effect through
complexation. In this regard, BLM speciation output for each of the silver concentrations tested showed that 90% of the silver was
as AgCl and AgCl2. High chloride levels have
been shown to protect O. mykiss against silver toxicity presumably by complexation to
form AgCl, thereby reducing the concentration of Ag+. However, the formation of AgCl
does not appear to substantially influence silver sensitivity for any other fish species tested
(Bielmyer et al. 2008). Therefore, it remains to
determine experimentally whether changes in
silver speciation related to changes in chloride concentration reduce silver toxicity to C.
decemmaculatus.
Water pH is another water quality parameter
that would affect silver toxicity through speciation. During the present toxicity test, mean
water pH was 8.3. In this regard, Erickson et
al. (1998) reported a reduction in silver toxicity
with increasing pH. The 96-h LC50 at pH 8.6
was three times higher than at pH 7.2. In the
present toxicity test, higher 96-h LC50 for C.
decemmaculatus may reflect some protective
effect against silver toxicity exerted by the elevated mean water pH.
Silver BLM LC50 prediction for P. promelas in
Pilcomayo River water did not agree well with
the LC50 calculated for C. decemmaculatus.
Bielmyer et al. (2007) in their silver BLM validation study found that only 50% of the LC50
predicted values for P. promelas were accurate within a factor of two. Higher 96-h LC50
for C. decemmaculatus may show that this
species is less sensitive to silver toxicity than
other fish species. However, it must be taken
into account that almost all of the studies previously cited on silver toxicity to P. promelas
worked with younger life-stages of the species which are known to be more sensitive to
pollutants. In this regard, Bielmyer et al. (2007)
reported higher 96-h LC50 for P. promelas as
fish aged and Grosell and co-authors (2002)

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Acta Toxicol. Argent. (2013) 21 (2): 93-101

reported higher sodium turnover in smaller

animals compared with larger animals in fresh
water. Bianchini et al. (2002) studied LC50 values based on total silver as a function of the
body mass. According to their results, log LC50
progressively increased with log body weight,
independent of the species considered. For
the mean body weight of the animals tested
in the present contribution, the relationship
found by these authors predicted a lower LC50
(0.45 g Ag/L) compared to the value calculated for C. decemmaculatus (140 g Ag/L).
This difference may be another indicator of a
lower sensitivity of C. decemmaculatus to silver toxicity.
In the present contribution, a clear pattern of
mortality was observed. Most of the deaths
occurred within the first five hours. Due to the
fact that silver precipitation was observed, is
possible that fish were in contact with higher
silver concentrations during the first minutes to
hours of the toxicity test. We did not measure
silver accumulation on the gills but this pattern
of mortality would correspond to a peak and
decline in gill silver accumulation observed in
static toxicity tests. Nebeker et al. (1983); Erickson et al. (1998) and Morgan et al. (2004)
attributed this pattern of silver accumulation to
changes in silver bioavailability through complexation with the increasing organic matter. In
the present contribution, silver concentrations
tested were higher compared to other toxicity
tests performed, therefore, changes in silver
bioavailability may have occurred mainly by
silver precipitation.

and mortality. A cross-fish-species extrapolation of the P. promelas Ag BLM was not valid in
Pilcomayo River water and experimental conditions of this toxicity test.
Acknowledgements
This research was supported by grants of the University of
Buenos Aires. The authors want to thank Subsecretara de Recursos Hdricos-Argentina (SSRH) who kindly performed water sampling and monitoring operations at Misin La Paz. and
provided Pilcomayo River water quality data. The authors also
want to thank Mrs. Amalia Gonzlez for the artwork and Dr. Sergio Gomz and Dr. Jimena Gonzlez Naya for useful technical
suggestions.

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CONCLUSIONS
C. decemmaculatus higher Ag 96-h LC50 may
reflect that this species is less sensitive to silver than other fish species. The mortality pattern observed suggests that toxicity can be
attributed to the initial dissolved silver concentrations. The ameliorative effect of water
hardness on silver toxicity is not clear; however, Pilcomayo River water elevated hardness
may have exerted some protective effect. Dissolved organic carbon contribution in reducing silver bioavailabity was negligible. Elevated mean water pH may have exerted a major
protective effect. More research is needed to
determine the effects of high sodium and chloride concentrations on silver toxicity to C. decemmaculatus. The mortality pattern observed
in this toxicity test may lend some support to
a relationship between gill silver accumulation

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